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Old 05-16-2017, 08:19 AM   #1
RickC7
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Location: Baton Rouge, Louisiana

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Default TruSeq Stranded Total RNA + NEB Barcodes??

Looking to get more index possibilities out of a TruSeq Stranded Total RNA LT kit. NEB Ultra directional workflow looks the same and NEB will sell the extra barcode kit alone. As far as I can tell, it looks like the NEB index adapters will work with the TruSeq kit, since both use dA-tailing... NEB #E7335, #E7500. Anyone have experience with this?

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Old 05-17-2017, 04:39 AM   #2
MU Core
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Keep in mind you'll require treatment of the adapter ends with the USER enzyme to free the circularized ends for downstream sequencing chemistry. This is a step you would have to incorporate into the TruSeq workflow. Not sure how easy that would be with the master mixes, etc. of the TruSeq kit.
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Old 05-17-2017, 05:01 AM   #3
RickC7
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Yes, I found a couple of caveats and work-arounds, but thankfully I won't need to employ any customization as Illumina gratefully offered to send me extra barcodes for this instance.
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